Daniel:Notebook/PosSequencing/2017-5-17: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Djacobse
>Djacobse
Line 9: Line 9:
'''Alkaline Lysis Solution'''
'''Alkaline Lysis Solution'''


*80 ul Nuclease free H<sub>2</sub>O.
*97 ul Nuclease free H<sub>2</sub>O.
*8 ul 5 M KOH.
*1 ul 100 mM KOH
*10 ul 1 M DTT.
*2 ul 0.5 M EDTA
*2 ul 0.5 M EDTA



Revision as of 18:37, 17 May 2017

Bead Binding Test 3

Back to Calendar

Avidin-Biotin Bonds Paper

Trying to test the bead binding and denaturation on the beads again. This time we'll use high pH as the denaturant. From the paper above it says the streptavidin bond is stable to about pH 11, and according to this website DNA denaturation occurs at pH>10. I will use the alkaline lysis solution from Andrew's notes

Alkaline Lysis Solution

  • 97 ul Nuclease free H2O.
  • 1 ul 100 mM KOH
  • 2 ul 0.5 M EDTA

Neutralization Solution

  • 3 mL H20
  • 3 mL 2M Tris-HCl
  • 4 mL 1N HCl

Protocol